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Differential partitioning by disorder-mediated condensates

ABSTRACT: Samples 1012398_F1-F8: Using recombinant purified mCherry-tagged FUS-IDR we reconstituted condensates in a soluble nuclear extract isolated from U2OS cells. The extract was fractionated by centrifugation into a supernatant and pellet. Labels 126, 127, 128 are supernatant replicates and labels 129, 130, 131 are pellet replicates. Samples 994350_F1-8: Using recombinant purified mEGFP-tagged MED1-IDR we reconstituted condensates in a soluble nuclear extract isolated from U2OS cells. The extract was fractionated by centrifugation into a supernatant and pellet. Labels 126, 127, 128 are supernatant replicates and labels 129, 130, 131 are pellet replicates. For the proximity biotinylation method, we used SILAC (Lys-6, Arg-6) in the media of cells expressing the control while both FUS-IDR and MED1-IDR cells were grown in natural isotope conditions. Cells were subject to the biotinylation protocol followed by nuclear extract preparation. Light and heavy extracts were mixed at a 1:1 ratio followed by streptavidin pulldown, stringent washing, and elution. Two independent replicates of SILAC data were collected for both FUS-IDR (QEX2_1011001 and ECL1_1073837) and MED1-IDR (QEX2_996337 and ECL1_1078014).

INSTRUMENT(S): Orbitrap Eclipse, Orbitrap Fusion Lumos, Q Exactive HF

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Benjamin Sabari

PROVIDER: MSV000093520 | MassIVE | Tue Nov 28 08:26:00 GMT 2023

REPOSITORIES: MassIVE

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